Clonal barcoding with qPCR detection enables live cell functional analyses for cancer research
Qiuchen Guo1,2, Milos Spasic1,2, Adam G Maynard2
1Division of Hematology, Department of Medicine, Brigham and Women's Hospital, Boston, MA, 02115, USA.
Nature Communications
|July 5, 2022
Summary
SunCatcher enables live single-cell retrieval and functional analysis using clonal molecular barcoding. This method tracks individual cells longitudinally, revealing diverse clonal evolution and metastasis in cancer research.
Area of Science:
- Cell Biology
- Molecular Biology
- Cancer Research
Background:
- Current single-cell analysis methods lack live cell retrieval capabilities.
- Studying eliminated cells is crucial for understanding experimental outcomes and disease progression.
Purpose of the Study:
- To develop a method for longitudinal tracking and live cell functional analysis.
- To enable the study of individual cell clones within complex populations.
Main Methods:
- Developed SunCatcher, a clonal molecular barcoding method.
- Generated single cell-derived clonal populations infected with unique molecular barcodes.
- Utilized qPCR and next-generation sequencing for barcode identification and quantification in vitro and in vivo.
Main Results:
- SunCatcher enables longitudinal tracking and functional analysis of live single-cell clones.
- Applied to breast cancer cell lines, revealing phenotypic and functional diversity among individual clones.
- Identified and quantified early spontaneous metastases.
- Heterogeneous barcoded clone pools recapitulated parental cell line proliferation and tumor progression.
Conclusions:
- SunCatcher offers a rapid and sensitive approach for studying live single-cell clones and clonal evolution.
- Facilitates functional analyses of individual cells and their contributions to disease progression.
- Provides critical insights into cellular heterogeneity and metastasis in cancer.
